Substituted sulfonamides and ureas useful for inhibiting kinase activity

ABSTRACT

Sulfonamide and urea compounds having an inhibitory effect on Src kinase including enantiomers, stereoisomers and tautomers thereof, as well as pharmaceutically acceptable salts or solvates of said compound, said compound having the general structure shown in Formula II are disclosed and claimed:

This application is a division of U.S. application Ser. No. 10/835,630,filed Apr. 30, 2004, now allowed, which is a division of U.S.application Ser. No. 10/191,718, filed Jul. 9, 2002, now U.S. Pat. No.6,777,577, issued Aug. 17, 2004, which claims the benefit of U.S.Provisional Application No. 60/304,020, filed Jul. 9, 2001.

FIELD OF THE INVENTION

The present invention discloses novel substituted amide compounds,specifically sulfonamides, and urea compounds having enzyme inhibitingproperties, especially for inhibiting protein tyrosine kinases.

BACKGROUND OF THE INVENTION

The novel compounds of the present invention may have generaltherapeutic value for the treatment of such diseases as cancer,including bone, colon or breast cancer; immunodeficiency disorders anddiabetes; atherosclerosis, osteoporosis, leukemia and other conditionssuch as coronary heart disease, congestive heart failure, renal failureand diseases of the central nervous system where the compounds exert abeneficial effect. The inventive compounds have been found to inhibitthe Src protein tyrosine kinase, a member of the Src family.

The Src family consists of nine members—Src. Yes, Fgr, Yrk. Fyn. Lyn,Hck, Lck and Blk—which share the same domain structure. The N-terminal,unique domain contains a myristylation site and frequently apalmitoylation site. It is followed by the regulatory SH3 and SH2domains, a catalytic domain that is bilobal and has its active sitewedged between the two lobes, and a C-terminal regulatory tail thatcontains the hallmark regulatory tyrosine residue (Tyr527 in Src).Kinase activity is reduced when the latter is phosphorylated and boundto the SH2 domain. The SH2 and SH3 domains bind phosphotyrosyl andproline-rich peptides, respectively: through these interactions, theyparticipate in intra- and intermolecular regulation of kinase activity,as well as localization and substrate recognition.

There is a wealth of evidence that tyrosine phosphorylation plays acrucial role in many cell regulatory processes. Fahad Al-Obeidi et al.,Biopolymers (Peptide Science) 47, 197-223 (1998). Researchers have foundthat functional perturbation of the kinases results in many diseases.Thus, there has been a great deal of effort applied in attempts todevelop potent and selective inhibitors for these enzymes.

The Src protein tyrosine kinase plays a role in osteoporosis and otherbone diseases. Osteoporosis is defined as a systemic skeletal diseasewhich is characterized by low bone mass and microarchitecturaldeterioration of bone tissue resulting in an increase in bone fragilityand susceptibility to fracture, W. A. Peck, et al., Am. J. Med., 94,646, (1993) Conference Report. It is estimated that osteoporosis causes1.5 million fractures annually with a total medical cost of $13.8billion. National Osteoporosis Foundation, August, 1997. The mosttypical sites of such fractures are the hip, spine, wrist, and ribs. Itis also estimated that one out of every two women and one in eight menwill have an osteoporosis related fracture in their lifetime.Osteoporosis is most commonly associated with postmenopause andage-related bone tissue loss. In addition, osteoporosis can occursecondarily to various drugs and diseases such as corticosteroids,anticonvulsants, alcohol, malabsorption syndromes, primary biliarycirrhosis, myeloma, thalassemia, thyrtoxicosis, Cushing's syndrome,Turner's syndrome, and primary hyperparathyroidism. Drugs used in thetreatment of osteoporosis are generally classified as antiresorptive orformation stimulating. In normal bone tissue, there is a balance betweenbone formation by osteoblasts and bone resorption by osteoclasts. Whenthe balance of this ongoing process is upset, bone resorption can exceedbone formation resulting in quantitative bone loss. Most of thetreatments have involved those that act through inhibition of boneresorption, such as calcium supplements, estrogen, calcitonin, andvitamin D, L. Riggs, West. J. Med., 154, 63 (1991).

Examples of treatments which act though stimulation of bone formationare sodium fluoride, low intermittent dosage of parathyroid hormone, M.Missbach, et al., Rech. Chimie Med., July, 1997, London.

Several reports have disclosed compelling evidence that the proteintyrosine kinase (PTK)p60c-Src (sometimes referred to as c-Src) plays acritical role in osteoclastic function, M. Missbach, et al., ibid. Itwas reported that, in vitro, kinase inhibitors of c-Src are capable ofreducing osteoclastic bone resorption, Ibid. Osteoclasts are bone marrowcells that are responsible for breaking down or remodeling bone. Once anosteoclast comes into contact with the bone surface, it adheres tightlyto the bone, flattens out, and begins the process of secreting materialswhich results in dissolution of the bone. This fundamental action ofosteoclasts is dependent on Src kinase. In this case it is clear that atleast one of the roles for Src kinase is in the regulation ofcytoskeletal changes involved in establishing the close bone cellinterface and in polarizing cellular secretion toward the bone surface.Thus, animals genetically engineered to lack Src kinase showabnormalities that indicate a general inability to resorb bone.

In addition, osteoclasts derived from these animals are neither able toflatten on bone, nor are they able to dissolve it. Consistent with theseresults, small molecule inhibitors of Src kinase have been shown to beuseful in countering bone loss in animal models of osteoporosis, such asIL-1-induced hypercalcemia, and bone loss in ovariectomized rats. Srckinase inhibitors would be useful for the treatment of disorders markedby inappropriate bone resorption like osteoporosis.

SUMMARY OF THE INVENTION

The present invention provides novel carboxamides, sulfonamides andureas having inhibitory activity against osteoporosis and related bonetissue loss.

In one embodiment, this invention provides novel carboxamide compoundshaving such desirable therapeutically inhibitory activity. The inventivecarboxamides have the general structure shown in Formula I, includingenantiomers, stereoisomers and tautomers thereof, or pharmaceuticallyacceptable salts or solvates of said compound, said compound having thegeneral structure shown in Formula I:

-   -   wherein:    -   m is an integer from 0 to 4;    -   n is an integer from 1 to 6;    -   R′ is a C₁-C₄ alkyl;    -   R₁ is:

-   -   R₃ is selected from the group consisting of H, C₁-C₄ straight        chain alkyl and C₁-C₄ branched alkyl;    -   R₂ is selected from the group consisting of        —(CH₂)_(p)—NH—C(═NH)NH₂; —(CH₂)_(p)—R₄; and —(CH₂)_(q)—Ar₁,        wherein p is an integer from 1 to 4; q is 0 or 1; R₄ is C₅-C₇        cycloalkyl; and Ar₁ is selected from the group consisting of:

-   -   wherein R₅ is —NH₂ or phenyl.

In another embodiment, this invention provides novel substituted ureaand sulfonamide compounds having inhibitory activity againstosteoporosis and related bone tissue loss. The inventive compounds havethe general structure shown in Formula II, including enantiomers,stereoisomers and tautomers thereof, as well as its pharmaceuticallyacceptable salts or solvates:

-   -   wherein R₁ is selected from the group consisting of H, straight        chain C₁-C₆ alkyl; branched C₁-C₆ alkyl; —(CH₂)_(p)—Ar₁; and        —(CH₂)_(p)—R₄, wherein p is 1 or 2;    -   Ar₁ is phenyl or naphthyl optionally substituted with a straight        chain or branched C₁-C₆ alkyl group; and    -   R₄ is C₅-C₇ cycloalkyl;    -   R₂ is selected from the group consisting of:

-   -   R₅ is selected from the group consisting of H, straight chain        C₁-C₆ alkyl; branched C₁-C₆ alkyl;    -   R₃ is —(CH₂)_(q)—Ar₂ or —(CH═CH)-Phenyl, wherein q is an integer        from 0 to 4; and Ar₂ is selected from the group consisting of:

-   -   X is:

-   -   Y is selected from the group consisting of:

-   -   with the proviso that when Y is any of the moieties:

then R₁ is H.When used herein, unless otherwise defined, the following terms have thegiven meanings:

alkyl (including the alkyl portions of lower alkoxy)—represents astraight or branched, saturated hydrocarbon chain having from 1 to 10carbon atoms, preferably from 1 to 6;

aryl—represents a carbocyclic group having from 6 to 14 carbon atoms andhaving at least one benzenoid ring, with all available substitutablearomatic carbon atoms of the carbocyclic group being intended aspossible points of attachment. Preferred aryl groups include 1-naphthyl,2-naphthyl and indanyl, and especially phenyl and substituted phenyl;

aralkyl—represents a moiety containing an aryl group linked vial a loweralkyl;

alkylaryl—represents a moiety containing a lower alkyl linked via anaryl group;

cycloalkyl—represents a saturated carbocyclic ring having from 3 to 8carbon atoms, preferably 5 or 6, optionally substituted.

heterocyclic—represents, in addition to the heteroaryl groups definedbelow, saturated and unsaturated cyclic organic groups having at leastone O, S and/or N atom interrupting a carbocyclic ling structure thatconsists of one ring or two fused rings, wherein each ring is 5-, 6- or7-membered and may or may not have double bonds that lack delocalized pielectrons, which ring structure has from 2 to 8, preferably from 3 to 6carbon atoms, e.g., 2- or 3-piperidinyl, 2- or 3-piperazinyl, 2- or3-morpholinyl, or 2- or 3-thiomorpholinyl;

halogen—represents fluorine, chlorine, bromine and iodine;

heteroaryl—represents a cyclic organic group having at least one O, Sand/or N atom interrupting a carbocyclic ring structure and having asufficient number of delocalized pi electrons to provide aromaticcharacter, with the aromatic heterocyclic group having from 2 to 14,preferably 4 or 5 carbon atoms, e.g., 2-, 3- or 4-pyridyl, 2- or3-furyl, 2- or 3-thienyl, 2-, 4- or 5-thiazolyl, 2- or 4-imidazolyl, 2-,4- or 5-pyrimidinyl, 2-pyrazinyl, or 3- or 4-pyridazinyl, etc. Preferredheteroaryl groups are 2-, 3- and 4-pyridyl; such heteroaryl groups mayalso be optionally substituted.

The term “pharmaceutically acceptable salt” is a non-toxic organic orinorganic acid addition salt of the base compounds represented byFormulas I and II.

Included within the scope of the present invention are the individualstereoisomers, diastereomers and geometric isomers of formula (I) and(II), and enantiomers thereof. The term “stereoisomers” is a generalterm for all isomers of individual molecules that differ only in theorientation of their atoms in space. It includes geometric (cis/trans)isomers, and isomers of compounds with more than one chiral center thatare not mirror images of one another (diastereomers). The term“enantiomer” or “enantiomeric” refers to a molecule that isnonsuperimposable on its mirror image and hence optically active whereinthe enantiomer rotates the plane of polarized light in one direction andits mirror image rotates the plane of polarized light in the oppositedirection. The term “racemic mixture” or “racemic modification” refersto a mixture of equal parts of enantiomers and which is opticallyinactive. As used herein the prefixes “(+)” and “(−)” are employed todesignate the sign of rotation of the plane of polarized light by thecompound, with (+) meaning the compound is dextrorotatory and (−)meaning the compound is levorotatory. For amino-acids, the designationsL/D, or R/S can be used as described in IUPAC-IUB Joint Commission onBiochemical Nomenclature, Eur. J. Biochem. 138, 9-37 (1984).

A further feature of the invention is pharmaceutical compositionscontaining as active ingredient a compound of Formula I (or its salt,solvate or isomers) or Formula II (or its salt, solvate or isomers)together with a pharmaceutically acceptable carrier or excipient.

The invention also provides methods for administering to a patientsuffering from one or more of the aforesaid diseases a therapeuticallyeffective inhibitory amount of a compound of Formula I or Formula II, orpharmaceutical compositions comprising a compound of Formula I orFormula II.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment, the present invention provides novel compounds ofFormula I or Formula II shown above, where the various symbols are asdefined. Representative amide compounds of the invention which exhibitexcellent Src kinase inhibitory activity belonging to Formula I arelisted below by names and structure.

Names and Structural Formulas

-   1.    N-[4-Amidinobenzoyl]-N-[3-phenoxybenzyl]-3-(4-biphenyl)-alanyl-glycyl-amide

IUPAC Name:

-   -   Alpha-[[4-(aminoiminomethyl)        benzoyl][(3-phenoxy-phenyl)methyl]amino]-N-(2-amino-2-oxoethyl)-1,1′-biphenyl-4-propanamide

Structure:

-   2.    N-[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-cyclohexylalanyl-glycyl-amide

IUPAC Name:

-   -   3-(aminoiminomethyl)-N-[1-[[(2-amino-2-oxoethyl)amino]carbonyl]-2-cyclohexylethyl]-N-[[3-[4-(1,1-dimethylethyl)phenoxy]phenyl]methyl]benzamide

Structure:

-   3.    N-[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-4-aminophenylalanyl-glycyl-amide

IUPAC Name:

-   -   4-amino-alpha-[[3-(aminoiminomethyl)benzoyl][[3-[4-(1,1-dimethylethyl)phenoxy]phenyl]methyl]amino]-N-(2-amino-2-oxoethyl)benzenepropanamide

Structure:

-   4.    N-[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-1-naphthylalanyl-glycyl-amide

IUPAC Name:

-   -   4-amino-alpha-[[3-(aminoiminomethyl)benzoyl][[3-[4-(1,1-dimethylethyl)phenoxy]phenyl]methyl]amino]-N-(2-amino-2-oxoethyl)-1-naphthalenepropanamide

Structure:

-   5.    N-[[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-arginyl-glycyl-amide

IUPAC Name

-   -   3-(aminoiminomethyl)-N-[4-[(aminoiminomethyl)        amino]-1-[[(2-amino-2-oxoethyl)amino]carbonyl]butyl]-N-[[3-[4-(1,1-dimethylethyl)phenoxy]phenyl]methyl]benzamide

Structure:

-   6.    N-[4-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-tryptanyl-glycyl-amide

IUPAC Name:

-   -   4-amino-alpha-[[4-(aminoiminomethyl)benzoyl][[3-[4-(1,1-dimethylethyl)phenoxy]phenyl]methyl]amino]-N-(2-amino-2-oxoethyl)1H-indole-3-propanamide

Structure:

-   7.    N-[4-Amidinobenzoyl]-N-[4-biphenylmethyl]-3-(4-biphenyl)alanyl-glycyl-amide

IUPAC Name:

-   -   Alpha-[[4-(aminoiminomethyl)benzoyl][[[1,1′-biphenyl]-4-yl]methyl]amino]-N-(2-amino-2-oxoethyl)-1,1′biphenyl-4-propanamide

Structure:

Representative urea compounds of Formula II of the invention whichexhibit excellent Src kinase inhibitory activity are listed below bynames and structure.

-   8.    4-Cyclohexyl-1-[[2-(4-phenylbutanoyl)amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl]piperazine

IUPAC Name:

-   -   Alpha-[[[[4-(4-cyclohexyl-1-piperazinyl)-3-[(1-oxo-4-phenylbutyl)amino]phenyl]amino]carbonyl]amino]-2-naphthalenepropanamide

Structure:

-   9.    4-Cyclohexyl-1-[[2-cinnamoylamino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl]piperazine

IUPAC Name:

-   -   Alpha-[[[[4-(4-cyclohexyl-1-piperazinyl)-3-[(1-oxo-3-phenyl-2-propenyl)amino]phenyl]amino]carbonyl]amino]-2-naphthalenepropanamide

Structure:

-   10. Common Name:    -   4-Cyclohexyl-1-[[2-cinnamoylamino]-4-[(1-aminocarbonyl-3-phenyl)propylamino]carbonylaminophenyl]piperazine

IUPAC Name:

-   -   Alpha-[[[[4-(4-cyclohexyl-1-piperazinyl)-3-[(1-oxo-3-phenyl-2-propenyl)amino]phenyl]amino]carbonyl]amino]benzenebutanamide

Structure:

-   11.    4-Cyclohexyl-1-[[2-(4-phenylbutanoyl)amino]-4-[(1-aminocarbonyl-3-phenyl)propylamino]carbonylaminophenyl]piperazine

IUPAC Name

-   -   Alpha-[[[[4-(4-cyclohexyl-1-piperazinyl)-3-[(1-oxo-4-phenylbutl)amino]phenyl]amino]carbonyl]amino]benzenebutanamide

Structure:

-   12.    4-Cyclohexyl-1-[[2-cinnamoylamino]-4-[(1-aminocarbonyl-2-cyclohexylethylamino)carbonylaminophenyl]piperazine

IUPAC Name:

-   -   2-[alpha-[[[[4-(4-cyclohexyl-1-piperazinyl)-3-[(1-oxo-3-phenyl-2-propenyl)amino]phenyl        amino]carbonyl]amino]]3-(cyclohexyl)propanamide

Structure:

-   13. 4-(Piperidin-4-yl)carbonyl-1-[[2-(4-phenylbutanoyl)    amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]-carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   2-[alpha-[[[[4-[hexahydro-4-(4-piperidinylcarbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-oxo-5-phenylpentyl)amino]phenyl]amino]carbonyl]amino]]-3-[naphth-2-yl]propanamide

Structure:

-   14. 4-(Piperidin-4-yl)carbonyl-1-[[2-(2-benzofuranoyl)    amino]-2-(2-naphthyl)ethylamino]carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   2-[alpha-[[[[4-[hexahydro-4-(4-piperidinylcarbonyl)-1H-1,4-diazepin-1-YL]-3-[(1-oxo-1-benzofuran-2-yl)        amino]phenyl]amino]carbonyl]amino]]-3-(naphth-2-yl)-propanamide

Structure:

-   15. 4-(Piperidin-4-yl)carbonyl-1-[[2-(2-benzofuranoyl)    amino]-4-[1-aminocarbonyl-2-cyclohexylethylamino]carbonylamino    phenyl]homopiperazine

IUPAC Name:

-   -   2-[alpha-[[[[4-[hexahydro-4-(4-piperidinylcarbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-oxo-1-benzofuran-2-yl)        amino]phenyl]amino]carbonyl]amino]]-3-cyclohexyl-propanamide

Structure:

-   16. 4-(Piperidin-4-yl)carbonyl-1-[2-[(2-benzofuranoyl)    amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   4-[alpha-[4-[[[[[4-[hexahydro-4-(4-piperidinyl-carbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-oxo-1-benzofuran-2-yl)amino]phenyl]amino]carbonyl]amino]methyl]]-cyclohex-1-ylformamide

Structure:

-   17. 4-(Methylaminomethyl)carbonyl-1-[2-[(2-benzo-furanoyl)    amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   4-[alpha-[4-[[[[[4-[hexahydro-4-(4-[[[methyl]amino]methyl]carbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-OXO-1-benzofuran-2-yl)amino]phenyl]amino]carbonyl]amino]methyl]]cyclohex-1-ylformamide

Structure:

-   18.    4-(Pyrrolidin-2-yl)carbonyl-1-[2-[(2-benzo-furanoyl)amino]-4-[[(4-aminocarbonyl)cyclohexyl-methyl-amino]carbonylamino]phenyl]homopiperazine

IUPAC Name:

-   -   4-[alpha-[[[[4-[hexahydro-4-(2-pyrrolidinyl-carbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-oxo-1-benzofuran-2-yl)amino]phenyl]amino]carbonyl]amino]methyl]]-cyclohex-1-ylformamide

Structure:

-   19.    4-(Piperidin-1-yl)-1-[2-[(2-benzofuranoyl)amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylaminophenylpiperidine

IUPAC Name:

-   -   4-[alpha-[4-[[[[[4-[4-(piperidin-1-yl)-1-piperidinyl]-3-[(1-oxo-1-benzofuran-2-yl)amino]phenyl]amino]carbonyl]amino]methyl]]-cyclohex-1-ylformamide

Structure:

-   20. 4-(Piperidin-4-yl)carbonyl-1-[[2-(4-phenylbutanoyl)    amino]-4-[1-aminocarbonyl-2-cyclohexylethylamino]carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   N-[5-[[[1-(carboxy)-1-(cyclohexyl-methyl)]amino]-carbonyl]amino]-2-[hexahydro-4-(4-piperidinyl-carbonyl)-1H-1,4-diazepin-1-yl]phenyl]benzene-butanamide

Structure:

-   21. 4-(Piperidin-4-yl)carbonyl-1-[[2-(4-phenylbutanoyl)    amino]-4-[(1-aminocarbonyl-2-(naphth-2-yl))ethylamino]-carbonylaminophenyl]homopiperazine

IUPAC Name:

-   -   2-[alpha-[[[[4-[hexahydro-4-(4-piperidinylcarbonyl)-1H-1,4-diazepin-1-yl]-3-[(1-oxo-4-phenyl-butyl)        amino]phenyl]amino]carbonyl]amino]]-3-(naphth-2-yl)-propanamide

Structure:

Representative sulfonamide compounds of the invention which exhibitexcellent Src kinase inhibitory activity of the Formula II are listed bynames and structure below.

-   22.    N-(1-Aminocarbonyl-2-methylpropyl)-2-[(4-phenylmethyl)piperidin-1-yl]-5-[(2-pyrrolidinocarbonyl)amino]phenylsulfonamide

IUPAC Name:

-   -   Alpha-2-[[[[2-(4-benzyl)-1-piperidinyl]-5-[(pyrrolidin-2-yl)carbonylamino]phenyl]sulfonyl]amino]-3-methylbutanamide

Structure:

-   23.    N-(1-Aminocarbonyl-2-methylpropyl)-2-[(4-phenylmethyl)piperidin-1-yl]-5-[(2-piperdino-carbonyl)    amino]phenylsulfonamide

IUPAC Name:

-   -   Alpha-2-[[[[2-(4-benzyl)-1-piperidinyl]-5-[(piperidin-4-yl)carbonylamino]phenyl]sulfonyl]amino]-3-methylbutanamide

Structure:

-   24.    1-[2-[N-(2-Aminocarbonyl-3-methylbutyl)sulfonamido]-5-[2-cinnamoylamino]]phenyl-4-cyclohexylpiperazine

IUPAC Name:

-   -   Alpha-2-[[[2-(4-cyclohexyl-1-piperazinyl)-5-[(1-oxo-3-phenyl-2-propenyl)carbonylamino]phenyl]sulfonyl]amino]-3-methylbutanamide

Structure:

-   25.    N-[[(4-Aminocarbonyl)cyclohexylmethyl]amino]-[2-[(4-phenylmethyl)piperidin-1-yl]-5-[(2-pyrrolidino-carbonyl)amino]phenyl]sulfonamide

IUPAC Name:

-   -   4-[2-[[[[[2-[(4-benzyl)-1-piperidinyl]-5-[(pyrrolidin-2-yl)carbonylamino]phenyl]sulfonyl]amino]methyl]]-cyclohex-1-ylformamide

Structure:

The compounds of the invention may form pharmaceutically acceptablesalts with organic and inorganic acids. Examples of suitable acids forsuch salt formation are hydrochloric, sulfuric, phosphoric, acetic,citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic,maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic,salicyclic, 2-phenoxybenzoic, p-toluensulfonic acid, and sulfonic acidssuch as methane sulfonic acid and 2-hyroxyethane sulfonic acid and othermineral and carboxylic acids well known to those skilled in the art andacid metal salts such as sodium monohydrogen orthophosphate, andpotassium hydrogen sulfate. Such salts can exist in either a hydrated orsubstantially anhydrous form. The salts are prepared by contacting thefree base form with a sufficient amount of the desired acid to produce asalt in the conventional manner. The free base forms may be regeneratedby treating the salt with a suitable dilute aqueous base solution suchas dilute aqueous sodium hydroxide, potassium carbonate, ammonia andsodium bicarbonate. The free base forms differ from their correspondingsalt forms somewhat in certain physical properties, such as solubilityin polar solvents, but the salts are otherwise equivalent to theircorresponding free base forms for purposes of this invention.

Depending upon the substituents on the inventive compounds, one may beable to form salts with bases too. Thus, for example, if there arecarboxylic acid substituents in the molecule (e.g., compound 21 in thelist above), salts may be formed with inorganic as well as organic basessuch as, for example, NaOH, KOH, NH₄OH, tetraalkylammonium hydroxide,and the like.

As stated earlier, the invention includes tautomers, enantiomers andother stereoisomers of the compounds also. Such variations arecontemplated to be within the scope of the invention.

Another embodiment of the invention discloses a method of making thesubstituted carboxamides, ureas and sulfonamides disclosed above. Thecompounds may be prepared by several processes known in the art ofsynthetic organic chemistry. One useful method to prepare compounds ofFormula I is schematically illustrated below in connection with thecompound numbered 7 above. In general, this procedure is referred to asScheme A herein and involves: (a) bonding an amino acid to a suitablyfunctionalized polymer support; (b) coupling another suitablysubstituted amino acid thereto; (c) reacting the coupled structure withan aldehyde to form a Schiff base, which is then (d) reduced to thecorresponding amine; which, in turn, is converted to an amide by way ofreaction with an acid chloride for example, which product may be (e)converted to the thioamide; which, in turn, is (f) methylated and (g)converted to the amidino group. The product is then cleared from thesolid phase support as will be further appreciated from the followingdiscussion.

Scheme A is specifically illustrated with respect toN-[4-Amidinobenzoyl]-N-[4-biphenylmethyl]-3-(4-biphenyl)alanyl-glycyl-amide:

In Scheme A, all substituents, unless otherwise indicated, are aspreviously defined. The reagents and starting materials are readilyavailable to one of ordinary skill in the art or may be prepared byconventional methods. The starting material (1) in Scheme A is an aminofunctionalized solid phase material, which for the purposes of synthesiswas modified with linker molecule (formula III), which enables theproduct of the synthesis to be cleaved from the solid support (resin).Example of such linker is the Rink linker(p-[(R,S)-α-(9H-fluoren-9-yl)methoxyformamide]-2,4-dimethoxybenzyl]-phenoxyaceticacid (Bernatowicz et al., Tetrahedron Lett. 30, 4645 (1989)).Commercially available resins with the desired linker already attachedcan be used as well.

The Rink linker attachment to a suitable solid phase is carried out byreacting an amino functionalized solid support with acid moiety of thelinker molecule by standard peptide synthesis techniques well known inthe art to provide an amide linkage, as shown in Example 1. Suchreaction can be carried out using standard coupling procedures such as,for example, as described in Stewart and Young, Solid Phase PeptideSynthesis, 2^(nd) ed., Pierce Chemical Co., Rockford, Ill. (1984);Gross, Meienhofer, Udenfriend, Ed., The Pepticles: Aiialysis, Synthesis,Biology, Vol. 1, 2, 3, 5 and 9, Academic Press, New York, 1980-1987;Bodanszky, Peptide Chemistry: A Practical Textbook, Springer-Verlag, NewYork (1988); and Bodanszky, et al. The Practice of Peptide SynthesisSpringer-Verlag, New York (1984), the disclosures of which are herebyincorporated by reference. If a coupling reagent (activator) is needed,suitable coupling reagent may be selected from dicyclohexylcarbodiimide(DCC), diisopropylcarbodiimide (DIC),1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquioline (EEDQ),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI),n-propanephosphonic anhydride (PPA),N,N-bis(2-oxo-3-oxazolidinyl)amidophosphoryl chloride (BOP-CI),diphenylphosphoryl azide, (DPPA), Castro's reagent (BOP),2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium salts (HBTU),2,5-diphenyl-2,3-dihydro-3-oxo-4-hydroxythiophene dioxide (Steglich'sreagent' HOTDO) and 1,1′carbonyldiimidazole (CDI). The coupling reagentmay be used alone or in combination with additives such as4-dimethylaminopyridine (DMAP), N-hydroxybenzotriazole (HOBt),N-hydroxybenzotriazine (HOOBt), N-hydroxysuccinimide) HOSu) or2-hydroxypyridine. The coupling reactions can be performed in eithersolution (liquid phase) or solid phase.

As used herein, the term “solid phase support” is not limited to aspecific type of support. A large number of supports are available andare known to one of ordinary skill in the art. Solid phase supportsinclude silica gels, resins, derivatized plastic films, glass beads,cotton, plastic beads, alumina gels, polysaccharides and the like. Asuitable solid phase support may be selected on the basis of desired enduse and suitability for various synthetic protocols. For example, forpeptide synthesis, solid phase support may refer to resins such asp-methylbenzhydrylamine (pMBHA) resin (from Peptides International,Louisville, Ky.), polystyrene (e.g., PAM-resin available from BachemInc. (Torance, Calif., USA), poly (dimethylacrylamide)-grafted styreneco-divinyl-benzene (e.g., POLYHIPE® resin, available from Aminotech,Nepean, Ontario, Canada), polyamide resin (e.g. Spar-resin, availablefrom AdvancedChemtech, Louisville, Ky., USA), polystyrene resin graftedwith polyethylene glycol (available from TentaGel®, Rapp Polymere,Tubingen, Germany) polydimethylacrylamide resin (available fromMilligen/Biosearch, Burlington, Mass., USA), or Sepharose (availablefrom Pharmacia Corporation, Stockholm, Sweden).

The amino acid moiety may carry protecting groups prior to the couplingreaction. Examples of suitable protecting groups include the following:(1) acyl types such as formyl, trifluoracetyl, phthalyl, andp-toluenesulfonyl; (2) aromatic carbamate types such asbenzyloxycarbonyl (Cbz or Z) and substituted benzyloxy-carbonyls,1-(p-biphenyl)-1-methylethoxy-carbonyl, and9-fluorenylmethyloxy-carbonyl (Fmoc); (3) aliphatic carbamate types suchas tertbutyloxycarbonyl (Boc), ethoxycarbonyl,diisopropyl-methoxycarbonyl, and allyloxycarbonyl; (4) cyclic alkylcarbamate types such as cyclopentyloxycarbonyl and adamantyloxycarbonyl;(5) alkyl types such as triphenyl-methyl and benzyl; (6) trialkysilanesuch as trimethyl-silane; and (7) thiol containing types such asphenylthio-carbonyl and dithiasuccinoyl. The preferred protecting groupis either Boc or Fmoc.

If certain functional groups or side chains on the amino acid moietyneed to be protected during the coupling reaction to avoid formation ofundesired bond, suitable protecting groups that can be used for thatpurpose are listed in Greene, Protective Groups in Organic Chemistry,John Wiley & Sons, New York (1981) and The Peptides: Analysis,Synthesis, Biology, Vol. 3, Academic Press, New York (1981), thedisclosures of which are hereby incorporated by reference. Those skilledin the art will appreciate the fact that the selection and use ofappropriate protecting groups depend upon the overall structure of theamino acid compound and the presence of any other protecting groups onthat compound. The selection of such a protecting group may beespecially important if it should not be removed during the deprotectionof the other protecting group.

Suitable amino acids for the coupling reaction are listed in Table 1along with the symbol for each amino acid.

TABLE 1 AMINO ACID SYMBOL Alanine Ala or A Arginine Arg or R AsparaginesAsn or N Aspartic acid Asp or D Cysteine Cys or C Glutamine Gin or QGlutamic acid Glu or E Glycine Gly or G Histidine His or H IsoleucineIle or I Leucine Len or L Lysine Lys or K Methionine Met or MPhenylalanine Phe or F Proline Pro or P Serine Ser or S Threonine Thr orT Tryptophan Trp or W Tyrosine Tyr or Y Valine Val or V

More specifically, a solid phase support such as, for example, adeprotected RAM-PS resin is typically treated with 3 equivalents of theamino acid moiety and 3 equivalents of 1-hydroxybenzotriazole in asuitable organic solvent, such a N,N-dimethylformamide. Then 3equivalents of diisopropylcarbodiimide are added and the mixture shakenfor about 30 minutes to five hours. The amide that is produced can beisolated and purified by well known techniques or the crude material canbe carried on to deprotection as it is.

The amide produced in the above-noted step is deprotected underconditions which do not cleave the solid phase support from the growingcompound. Such conditions are well known in the art. Thus, when the Bocprotecting group is used, the methods of choice are trifluoroacetic acideither neat or in dichloromethane, or HCl in dioxane or ethyl acetate.The resulting ammonium salt is then neutralized either prior to thecoupling or in situ with basic solutions such as aqueous buffers, ortertiary amines in dichloromethane or dimethylformamide. When the Fmocprotecting group is used, the reagents of choice are piperidine orsubstituted piperidine in dimethylformamide, but any secondary amine oraqueous basic solutions can be used. The deprotection is carried outgenerally at a temperature of between about 0° C. and about roomtemperature. For example, the above-noted crude amide may be treatedwith 30% piperidine in N,N-dimethylformamide for about 20 minutes toabout one hour, following which the reaction mixture is filtered toprovide the deprotected compound.

To the deprotected compound on solid phase, a suitably amino-protectedcompound having free carboxylic function (for example, Fmoc-protectedbiphenylalanine in Scheme A) to form the solid phase linked product. Forexample, 1 equivalents of the deprotected compound may be combined with3 equivalents of Fmoc-Biphenylalanine and 3 equivalents of1-hydroxybenzo-triazole and a suitable activator (for example 3equivalents of DIC) in a suitable organic solvent, such asN,N-dimethylformamide. The formed biphenylalanine linked compound iscleaved of the Fmoc group and then reacted with a suitable aldehyde,such as, for example, 4-phenylbenzaldehyde, to yield the correspondingSchiff base. The Schiff base is then reduced, for example, with sodiumborohydride, sodium cyanoborohydride and the like, to form thecorresponding amine which is then converted to the amide by reactingwith, for example, an acid chloride, in this case, 4-cyanobenzoylchloride. The amide may be converted to the thioamide which ismethylated and then converted to the amidino group. The product is thencleaved of the solid phase support to yield compound 7.

The compounds of Formula II where X is an urea may be prepared asdescribed in Scheme B:

Scheme B may be explained with the synthesis of a compound of Formula IIwhere R₁ is 2-naphthylmethyl, R₂ is cyclohexylpiperazinyl, R₃ is3-phenylpropyl, X is —NH—CO—, Y is CONH2 and n is 1. That compound iscompound 8 identified above,4-Cyclohexyl-1-[[2-(4-phenylbutanoyl)amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl]piperazine

Thus, a solid phase support is coupled with a protected amino acid, inthis case, Fmoc-2-naphthylalanine in the presence of an activator suchas, for example, 1-hydroxybenzotriazole and DIC. It is then deprotectedand then reacted with 4-fluoro-3-nitrophenylisocyanate and thefluorinated product is then reacted with 4-cyclohexylpiperazine tointroduce the R₂ group. The nitro group is then reduced with stannouschloride to the amine which is converted to the 4-phenylbutyl amide byreacting with 4-phenylbutyric acid by activation with HOAt and DIC.Cleaving of the solid support yields the desired compound 8. Similarly,one synthesizes the other urea compounds by appropriate selection of theR₁, R₂ and R₃ substituted reactants.

Synthesis of a compound of Formula II where X is sulfonamide is similarto that shown in Scheme B except that in the step introducing thefluoronitrophenyl-isocyanate, the appropriate fluoronitrobenzenesulfonyl chloride is used. Thus, replacing the isocyanate in the abovedescription with 2-fluoro-5-nitrobenzene sulfonyl chloride would yieldthe desired sulfonamide compound.

Isolation of the compound at various stages of the reaction scheme maybe achieved after cleavage from solid support by standard techniquessuch as, for example, filtration, evaporation of solvent and the like.Purification of the product, intermediate and the like, may also beperformed by standard techniques such as recrystallization,distillation, sublimation, chromatography, conversion to a suitablederivative which may be recrystallized and converted back to thestarting compound, and the like. Such techniques are well known to thoseskilled in the art.

The thus prepared compounds may be analyzed for their composition andpurity as well as characterized by standard analytical techniques suchas, for example, elemental analysis, NMR, mass spectroscopy, and IRspectra.

In another embodiment, this invention provides pharmaceuticalcompositions comprising the above-described inventive compounds as anactive ingredient. The pharmaceutical compositions generallyadditionally comprise a pharmaceutically acceptable carrier diluent,excipient or carrier (collectively referred to herein as carriermaterials). Because of their therapeutic activity against osteoporosisand bone tissue loss, such pharmaceutical compositions possess utilityin treating those diseases.

In yet another embodiment, the present invention discloses methods forpreparing pharmaceutical compositions comprising the compounds ofFormula I or Formula II as an active ingredient. In the pharmaceuticalcompositions and methods of the present invention, the active ingredientor ingredients will generally be administered in admixture with suitablecarrier materials suitably selected with respect to the intended form ofadministration, i.e. oral tablets, capsules (either solid-filled,semi-solid filled or liquid filled), powders for constitution, oralgels, elixirs, dispersible granules, syrups, suspensions, and the like,and consistent with conventional pharmaceutical practices. For example,for oral administration in the form of tablets or capsules, the activedrug component may be combined with any oral non-toxic pharmaceuticallyacceptable inert carrier, such as lactose, starch, sucrose, cellulose,magnesium stearate, dicalcium phosphate, calcium sulfate, talc,mannitol, ethyl alcohol (liquid forms) and the like. Moreover, whendesired or needed, suitable binders, lubricants, disintegrating agentsand coloring agents may also be incorporated in the mixture. Powders andtablets may be comprised of from about 5 to about 95 percent inventivecomposition. Suitable binders include starch, gelatin, natural sugars,corn sweeteners, natural and synthetic gums such as acacia, sodiumalginate, carboxymethylcellulose, polyethylene glycol and waxes. Amongthe lubricants there may be mentioned for use in these dosage forms,boric acid, sodium benzoate, sodium acetate, sodium chloride, and thelike. Disintegrants include starch, methylcellulose, guar gum and thelike.

Sweetening and flavoring agents and preservatives may also be includedwhere appropriate. Some of the terms noted above, namely disintegrants,diluents, lubricants, binders and the like, are discussed in more detailbelow.

Additionally, the compositions of the present invention may beformulated in sustained release form to provide the rate controlledrelease of any one or more of the components or active ingredients tooptimize the therapeutic effects, i.e. antihistaminic activity and thelike. Suitable dosage forms for sustained release include layeredtablets containing layers of varying disintegration rates or controlledrelease polymeric matrices impregnated with the active components andshaped in tablet form or capsules containing such impregnated orencapsulated porous polymeric matrices.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injections or addition of sweeteners and pacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier such as inert compressed gas, e.g.nitrogen.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides such as cocoa butter is first melted, and theactive ingredient is dispersed homogeneously therein by stirring orsimilar mixing. The molten homogeneous mixture is then poured intoconvenient sized molds, allowed to cool and thereby solidify.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions may take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

Preferably the compound is administered orally.

Preferably, the pharmaceutical preparation is in a unit dosage form. Insuch form, the preparation is subdivided into suitably sized unit dosescontaining appropriate quantities of the active components, e.g., aneffective amount to achieve the desired purpose.

The quantity of the inventive active composition in a unit dose ofpreparation may be generally varied or adjusted from about 1.0 milligramto about 1,000 milligrams, preferably from about 1.0 to about 950milligrams, more preferably from about 1.0 to about 500 milligrams, andtypically from about 1 to about 250 milligrams, according to theparticular application. The actual dosage employed may be varieddepending upon the patient's age, sex, weight and severity of thecondition being treated. Such techniques are well known to those skilledin the art.

Generally, the human oral dosage form containing the active ingredientscan be administered 1 or 2 times per day. The amount and frequency ofthe administration will be regulated according to the judgment of theattending clinician. A generally recommended daily dosage regimen fororal administration may range from about 1.0 milligram to about 1,000milligrams per day, in single or divided doses.

The term “capsule” refers to a special container or enclosure made ofmethylcellulose, polyvinyl alcohols, or denatured gelatins or starch forholding or containing compositions comprising the active ingredients.Hard shell capsules are typically made of blends of relatively high gelstrength bone and pork skin gelatins. The capsule itself may containsmall amounts of dyes, opaquing agents, plasticizers and preservatives.

The term “tablet” refers to a compressed or molded solid dosage formcontaining the active ingredients with suitable diluents. The tablet canbe prepared by compression of mixtures or granulations obtained by wetgranulation, dry granulation or by compaction.

The term “oral gel” refers to the active ingredients dispersed orsolubilized in a hydrophilic semi-solid matrix.

The term “powders for constitution” refers to powder blends containingthe active ingredients and suitable diluents which can be suspended inwater or juices.

The term “diluent” refers to substances that usually make up the majorportion of the composition or dosage form. Suitable diluents includesugars such as lactose, sucrose, mannitol and sorbitol; starches derivedfrom wheat, con, rice and potato; and celluloses such asmicrocrystalline cellulose. The amount of diluent in the composition canrange from about 10 to about 90% by weight of the total composition,preferably from about 25 to about 75%, more preferably from about 30 toabout 60% by weight, even more preferably from about 12 to about 60%.

The term “disintegrant” refers to materials added to the composition tohelp it break apart (disintegrate) and release the medicaments. Suitabledisintegrants include starches; “cold water soluble” modified starchessuch as sodium carboxymethyl starch; natural and synthetic gums such aslocust bean, karaya, guar, tragacanth and agar; cellulose derivativessuch as methylcellulose and sodium carboxymethylcellulose;microcrystalline celluloses and cross-linked microcrystalline cellulosessuch as sodium croscarmellose; alginates such as alginic acid and sodiumalginate; clays such as bentonites; and effervescent mixtures. Theamount of disintegrant in the composition can range from about 2 toabout 15% by weight of the composition, more preferably from about 4 toabout 10% by weight.

The term “binder” refers to substances that bind or “glue” powderstogether and make them cohesive by forming granules, thus serving as the“adhesive” in the formulation. Binders add cohesive strength alreadyavailable in the diluent or bulking agent. Suitable binders includesugars such as sucrose; starches derived from wheat, corn rice andpotato; natural gums such as acacia, gelatin and tragacanth; derivativesof seaweed such as alginic acid, sodium alginate and ammonium calciumalginate; cellulosic materials such as methylcellulose and sodiumcarboxymethylcellulose and hydroxypropylmethylcellulose;polyvinylpyrrolidone; and inorganics such as magnesium aluminumsilicate. The amount of binder in the composition can range from about 2to about 20% by weight of the composition, more preferably from about 3to about 10% by weight, even more preferably from about 3 to about 6% byweight.

The term “lubricant” refers to a substance added to the dosage form toenable the tablet, granules, etc. after it has been compressed, torelease from the mold or die by reducing friction or wear. Suitablelubricants include metallic stearates such as magnesium stearate,calcium stearate or potassium stearate; stearic acid; high melting pointwaxes; and water soluble lubricants such as sodium chloride, sodiumbenzoate, sodium acetate, sodium oleate, polyethylene glycols andd'l-leucine. Lubricants are usually added at the very last step beforecompression, since they must be present on the surfaces of the granulesand in between them and the parts of the tablet press. The amount oflubricant in the composition can range from about 0.2 to about 5% byweight of the composition, preferably from about 0.5 to about 2%, morepreferably from about 0.3 to about 1.5% by weight.

The term “glident” refers to materials that prevent caking and improvethe flow characteristics of granulations, so that flow is smooth anduniform. Suitable glidents include silicon dioxide and talc. The amountof glident in the composition can range from about 0.1% to about 5% byweight of the total composition, preferably from about 0.5 to about 2%by weight.

The term “coloring agent” refers to excipients that provide colorationto the composition or the dosage form. Such excipients can include foodgrade dyes and food grade dyes adsorbed onto a suitable adsorbent suchas clay or aluminum oxide. The amount of the coloring agent can varyfrom about 0.1 to about 5% by weight of the composition, preferably fromabout 0.1 to about 1%.

The term “bioavailability” refers to the rate and extent to which theactive drug ingredient or therapeutic moiety is absorbed into thesystemic circulation from an administered dosage form as compared to astandard or control.

Conventional methods for preparing tablets are known. Such methodsinclude dry methods such as direct compression and compression ofgranulation produced by compaction, or wet methods or other specialprocedures. Conventional methods for making other forms foradministration such as, for example, capsules, suppositories and thelike are also well known.

Another embodiment of the invention discloses use of the pharmaceuticalcompositions disclosed above for treatment of diseases such as, forexample, osteoporosis and bone tissue loss.

It will be apparent to those skilled in the art that many modifications,variations and alterations to the present disclosure, both to materialsand methods, may be practiced. Such modifications, variations andalterations are intended to be within the spirit and scope of thepresent invention.

The following examples are being provided to further illustrate thepresent invention. They are for illustrative purposes only; the scope ofthe invention is not to be considered limited in any way thereby.

EXAMPLES

Unless otherwise stated, the following abbreviations have the statedmeanings in the Examples below: DCC=dicyclohexylcarbodiimide

-   NaBH(OAc)₃=sodium triacetoxyborohydride-   FMOC=9-fluorenylmethyloxycarbonyl-   DCE=1,2-dichloroethane-   DIEA=diisopropylethylamine-   Cha=cyclohexylalanine-   NaI(1)=1-naphthylalanine-   TEOF=triethylorthoformate-   TIPS=triisopropylsilane-   NaI(1)=1-naphthylalanine-   Bip=4-biphenylalanine-   Boc=tert.butyloxycarbonyl-   Pip=piperidine-   HOAc=acetic acid-   TFA=trifluoroacetic acid-   Py=pyridine-   DIC=diisopropylcarbodiimide-   MeOH=methanol-   NaBH₄=sodium borohydride-   NaBH₃CN=sodium cyanoborohydride-   p-TsOH=p-toluenesulfonic acid-   DMF: N,N-Dimethylformamide-   THF: Tetrahydrofuran-   DMSO: Dimethyl sulfoxide-   DCM: Dichloromethane which can also be refered to as methylene    chloride-   LAH: Lithium aluminum hydride-   HOAt: 1-Hydroxy-7-azabenzotriazole-   HOBt: 1-Hydroxybenzotriazole-   HRMS=High Resolution Mass Spectrometry-   HPLC=High Performance Liquid Chromatography-   NMR=nuclear magnetic resonance-   LRMS=Low Resolution Mass Spectrometry-   nM=nanomolar

Additionally, “kg” refers to kilograms; “g” refers to grams; “mg” refersto milligrams; μg” refers to micrograms; “m²/g” refers to square metersper gram and is used as a measurement of particle surface area; “mmol”refers to millimoles; “L” refers to liters; “mL” refers to milliliters;“μL” refers to microliters; “cm” refers to centimeters; “M” refers tomolar’ “mM” refers to millimolar; “μM” refers to micromolar; “nM” refersto nanomolar; “N” refers to normal; “ppm” refers to parts per million;“δ” refers to parts per million down field from tetramethylsilane; “°C.” refers to degrees Celsius; “° F.” refers to degrees Fahrenheit; “mmHg” refers to millimeters of mercury; “kPa” refers to kilopascals; “psi”refers to pounds per square inch; “rpm” refers to revolutions perminute; “bp” refers to boiling point; “mp” refers to melting point;“dec” refers to decomposition; “h” refers to hours; “min” refers tominutes; “sec” refers to seconds’ “R_(f)” refers to retention factor;and “R_(t)” refers to retention time.

Examples 1-7 pertain to synthesis of compounds of Formula I.

General Synthesis Procedures

Starting materials used in the synthesis were obtained from chemicalvendors such as Aldrich, Sigma, Fluka, Nova Biochem and AdvancedChemtech. During the synthesis, the functional groups of the amino acidderivatives used were protected by blocking groups to prevent sidereaction during the coupling steps. Examples of suitable protectinggroups and their use are described in The Peptides, supra, 1981, and invol. 9, Udenfriend and Meienhofer (eds.), 1987, which is incorporatedherein by reference.

General solid-phase peptide synthesis was used to produce the compoundsof the invention. Such methods are described, for example, by Stewardand Young, Solid Phase Peptide Synthesis (Freeman & Co., San Francisco,1969), which is incorporated herein by reference.

Unless indicated otherwise, peptides were synthesized onRAM™-Polystyrene Resin (Rapp Polymere, Tübingen, Germany). As analternative to this, acid sensitive linkerp-[(R,S)-α-[1-(9H-fluoren-9-yl)methoxyformamido]-2,4-dimethoxybenzyl]phenoxyaceticacid (Knorr Linker, Bernatowicz et. al, Tetr. Lett. 30 (1989) 4645,which is incorporated herein by reference) can be coupled to any aminofunctionalized the solid support or the desired compounds can besynthesized on polystyrene resin cross-linked with 1% divinylbenzenemodified with an acid sensitive linker (Rink resin) (Rink, Tetr. Lett.28 (1987) 3787; Sieber, Tetr. Lett. 28 (1987) 2107, each of which isincorporated herein by reference). Coupling was performed usingN,N′-diisopropylcarbodiimide (DIC) in the presence of an equivalentamount of HOBt. All couplings were done N,N-dimethylformamide (DMF) atroom temperature (RT). Completion of coupling was monitored by ninhydrintest. A second (double) coupling was performed where coupling in thefirst instance was incomplete.

Deprotection of the Fmoc group was accomplished using 50% piperidine inDMF for 2+15 min. The amount of Fmoc released was determined from theabsorbance at 302 nm of the solution after deprotection, volume ofwashes and weight of the resin used in the synthesis.

The compound resin was at the end of the synthesis washed successivelywith DMF and DCM and the peptide was then cleaved and deprotected by amixture TFA/TIPS (99/1) for 2 hours, unless specified otherwise. Theresin was washed with DCM and the DCM wash combined with the TFAreleasate. The solution was evaporated, the product was redissolved in amixture of water and acetonitrile and lyophylized.

The dried compound was subjected to HPLC purification using anappropriate gradient of 0.1% TFA in water and acetonitrile (ACN). Aftercollecting the peak containing the intended synthetic product, thesolution was lyophilized and the compound was subjected to anidentification process, which included electrospray mass spectrum (MS)and/or NMR to confirm that the correct compound was synthesized.

For HPLC analysis, a sample of the product was analyzed using BeckmanHPLC system (consisting of 126 Solvent Deliver System, 166 ProgrammableDetector Module 507e Autosampler, controlled by Data Station with GoldNouveau software) and YMC ODS-AM 4.6×250 mm column at 230 nm and flowrate 1 ml/min.

For product purification, a sample of crude lyophilized compound wasdissolved in a mixture of 0.1% aqueous TFA containing 10% to 50% ACN.The solution of the product was usually filtered through a syringeconnected to a 0.45 μm “ACRODISC” 13 CR PTFE (Gelman Sciences; Ann ArborMich.) filter. A proper volume of filtered compound solution wasinjected into a semi-preparative C18 column (YMC ODS-A column (20×250mm), YMC, Inc., Wilmington, N.C.). The flow rate of a gradient orisocratic mixture of 0.1% TFA buffer and ACN (HPLC grade) as an eluentwas maintained using a Beckman “SYSTEM GOLD” HPLC (Beckman, System Gold,Programmable Solvent Module 126 and Programmable Detector Module 166controlled by “SYSTEM GOLD” software). Elution of the compound wasmonitored by UV detection at 230 nm. After identifying the peakcorresponding to the compound under synthesis using MS, the compound wascollected, lyophilized and biologically tested. MS was performed using aVG Platform (Fisons Instruments) instrument in ES+ mode. For NMR,typically samples were measured in DMSO-d₆ (Aldrich) using a BrukerAvance DPX 300 instrument.

Example 1N-[4-Amidinobenzoyl]-N-[3-phenoxybenzyl]-3-(4-biphenyl)-alanyl-glycyl-amide

Following generally the procedure described above as Scheme A,polystyrene-RAM (substitution 0.74 mmol/g, 100-200 mesh, Rapp Polymere,Tubingen, Germany, 0.5 g) was washed with DMF and the Fmoc-protectinggroup cleaved by 50% solution of piperidine in DMF (twice 10 minutes, 5ml each). The resin was then washed by DMF. Fmoc-Gly-OH (3 eq) activatedwith DICIHOBt (3 eq each) in DMF (3 ml) was coupled to the resinovernight and the completion was checked by ninhydrin test. AfterFmoc-group deprotection, the resin-bound intermediate was reacted withFmoc-4-biphenyl-alanine (3 eq, in 3 ml DMF) activated with DIC/HOBt (3eq each) overnight. Fmoc group was deprotected as described above andthe resin was washed with DMF. Resin was washed with DCM and a solutionof 3-phenoxybenzaldehyde (7 eq) in 5 ml TEOF/DCM (4:1) was added and thereaction was carried out for 6 hours, the resin was washed with DCM (3times) and the formed Schiff base was reduced with 5 ml of solutionNaBH₃CN overnight. This was prepared by mixing 1M NaBH₃CN in THF(commercially available) with DCE/MeOH/AcOH (80:18:2) in ratio 1:4.After the reduction resin was washed with MeOH, DMF, 10% DIEA in DMF,DMF and DCE. The resin-bound amine was reacted with 5 eq of4-cyanobenzoyl chloride in 5 ml DCE with 5 eq DIEA overnight. Resin wasthen washed with DCE, DMF, with mixture pyridine/Et₃N (2:1) and treatedwith 8 ml of saturated solution of H₂S in Pyridine/Et₃N (2:1). After 5hours, the solution was removed and the procedure repeated. Afterovernight standing, the resin was washed with acetone. The resultingthioamide was converted to the thioimidate by reaction with methyliodidein acetone ((4 ml of 20% solution, overnight). The resin was washed withacetone and MeOH, and a solution of 20 eq of ammonium acetate inmethanol containing 20 eq of acetic acid was added and the kept at 50°C. for 3 hours. The resin was then washed with MeOH, DMF and DCM. Theproduct was cleaved by TFA(1% TIPS). The crude product was purified bypreparative HPLC. MS analysis: calculated 625.3 (M), found 626.2 (MH)+.

Example 2 Preparation of3-amidinobenzoyl-(3-(4-tert-butylphenoxy)benzyl)-cyclohexylylalanyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH, Fmoc-Cha-OH,3-(4-tert. Butylphenoxy) benzaldehyde and 3-cyanobenzoyl chlorideaccording to procedures described in Example 1. MS analysis: calculated611.4 (M), found 612.3 (MH)+.

Example 3 N-[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-4-aminophenylalanyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH,Fmoc-Phe(4-NH-Boc)-OH, 3-(4-tert. Butylphenoxy) benzaldehyde and3-cyanobenzoyl chloride according to procedures described in Example 1.MS analysis: calculated 620.3 (M), found 621.3 (MH)+.

Example 4 N-[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-1-naphthylalanyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH, Fmoc-Nal(1)-OH,3-(4-tert. Butylphenoxy) benzaldehyde and 3-cyanobenzoyl chlorideaccording to procedures described in Example 1. MS analysis: calculated655.3 (M), found 656.2 (MH)+.

Example 5 N—[3-Amidinobenzoyl]-N-[3-(4-tert-butylphenoxy)benzyl]-arginyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH, Fmoc-Arg(Boc)2-OH,3-(4-tert. Butylphenoxy) benzaldehyde and 3-cyanobenzoyl chlorideaccording to procedures described in Example 1. MS analysis: calculated614.3 (M), found 615.2 (MH)+.

Example 64-amidinobenzoyl-(3-(4-tert-butylphenoxy)phenoxybenzyl)-tryptanyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH, Fmoc-Trp(Boc)-OH,3-(4-tert. Butylphenoxy) benzaldehyde and 4-cyanobenzoyl chlorideaccording to procedures described in Example 1. MS analysis: calculated644.3 (M), found 645.2 (MH)+.

Example 7N-[4-Amidinobenzoyl]-N-[4-biphenylmethyl]-3-(4-biphenyl)alanyl-glycyl-amide

The title compound was synthesized using Fmoc-Gly-OH, Fmoc-Bip-OH,4-phenylbenzaldehyde and 4-cyanobenzoyl chloride according to proceduresdescribed in Example 1. MS analysis: calculated 609.3 (M), found 610.2(MH)⁺.

Examples 8-15 describe the synthesis of compounds of Formula II where Xis a urea moiety.

Example 84-Cyclohexyl-1-{[2-(4-phenylbutanoyl)amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl}piperazine

Following generally the procedure described above in connection withScheme B, commercial Polystyrene-RAM resin (0.74 mmol/g) (Rapp Polymere,Tubingen, Germany, 0.25 g) was slurried in dichloromethane, washed withDMF and treated for 30 minutes with a mixture of piperidine and DMF (1:1v/v). The resin was washed with DMF (5×), DCM (5×) and DMF (3×) and thencoupled with 0.5 mmol of Fmoc-(L)-2-naphthylalanine,1-hydroxybenzotriazole and diisopropyl-carbodiimide in 3 ml DMFovernight. The resin was washed with DMF (5×) and treated withpiperidine/DMF again for 30 minutes. After washing as described above,the coupling with 0.5 mmol of 4-fluoro-3-nitrophenylisocyanate in 2 mlDMF was carried out over night. The resin was washed with DMF (5×) andtreated with 3 ml of a 0.5 molar solution of 1-cyclohexylpiperazine inDMF for 3 hours at 600. After washing with DMF (10×), the nitro groupwas reduced by shaking the resin with 4 ml of a molar solution of tinchloride dihydrate in DMF for 24 hours. The resin was washed with DMF(5×), MeOH (5×), DCM (5×), DMF containing 5% of diisopropylethylamine(1×) and DMF (3×). The final coupling with 1 mmol of 4-phenyl butyricacid, 1-hydroxy-7-azabenzotriazole and diisopropylcarbodiimide in 3 mlDMF was performed over night. Following extensive washing of the resinwith DMF, methanol and DCM and subsequent drying, it was cleaved with 3ml of 95% trifluoroacetic acid. The TFA solution was evaporated and theresidue was combined with the washings of the resin with methanol.Evaporation yielded the crude title compound which was purified bypreparative HPLC using the standard acetonitrile/-water+0.1% TFAgradient and a Vydac C-18 column. The pure sample had a M+1 ion at 661.3in the mass spectrum and was homogenous by HPLC with a retention time of26.95 minutes.

Example 9 4-Cyclohexyl-1-{[2-cinnamoylamino]-4-r1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl}piperazine

This was prepared by the method of Example 8 using trans-cinnamic acidin the final coupling step to give the title compound with M+1 ion at645.3 and a retention time of 26.88 minutes.

Example 104-Cyclohexyl-1-{[2-cinnamoylamino]-4-[(1-aminocarbonyl-3-phenyl)propylamino]carbonylaminophenyl}piperazine

This compound was prepared by the method of Example 8 usingFmoc-homophenylalanine in the initial coupling step to give the titlecompound with M+1 ion at 609.3 and retention time of 25.78 minutes.

Example 114-Cyclohexyl-1-{[2-(4-phenylbutanoyl)amino]-4-[(1-aminocarbonyl-3-phenyl)propylamino]carbonylaminophenyl}piperazine

This compound was prepared by the method of Example 8 usingFmoc-homophenylalanine in the initial coupling step to give the titlecompound with M+1 at 625.3 and a retention time of 26 minutes.

Example 124-Cyclohexyl-1-{[2-cinnamoylamino]-4-[(1-aminocarbonyl-2-cyclohexyl)ethylamino]carbonylaminophenyl}piperazine

This compound was prepared by the method of Example 8 usingFmoc-cyclohexylalanine in the initial coupling step to give the titlecompound with M+1 ion at 601.3 and retention time of 26.72 minutes.

Example 134-(Piperidin-4-yl)carbonyl-1-{2-(4-phenylbutanoyl)amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl}homopiperazine

Following generally the procedure shown in Scheme B above, commercialPolystyrene-RAM resin (0.74 mmol/g) (Rapp Polymere, Tubingen, Germany,0.25 g) was slurried in dichloromethane, washed with DMF and treated for30 minutes with a mixture of piperidine and DMF (1:1 v/v). The resin waswashed with DMF (5×), DCM (5×) and DMF (3×) and then coupled with 0.5mmol of Fmoc-(L)-2-naphthylalanine, 1-hydroxybenzotriazole anddiisopropylcarbodiimide in 3 ml DMF over night. The resin was washedwith DMF (5×) and treated with piperidine/DMF again for 30 minutes.After washing as described above, the coupling with 0.5 mmol of4-fluoro-3-nitrophenylisocyanate in 2 ml DMF was carried out over night.The resin was washed with DMF (5×) and treated with 3 ml of a 0.5 molarsolution of homopiperazine in DMF for 2 hours at 60°. The resin waswashed with DMF (10×) and coupled with 0.5 mmol of Boc-isonipecoticacid, HOBt and DIC in 2.5 ml of DMF over night. The resin was washedwith DMF (10×) and reduced with 4 ml of a molar solution of tin chloridedihydrate in DMF for 24 hours. The resin was washed with DMF (5×), MeOH(5×), DCM (5×), DMF containing 5% of diisopropyl-ethylamine (1×) and DMF(3×). The final coupling with 1 mmol of phenylbutyric acid,1-hydroxy-7-azabenzotriazole and diisopropylcarbodiimide in 3 ml DMF wasperformed over night. Following extensive washing of the resin with DMF,methanol and DCM and subsequent drying, it was cleaved with 3 ml of 95%trifluoroacetic acid. The TFA solution was evaporated and the residuewas combined with the washings of the resin with methanol. Evaporationyielded the crude title compound which was purified by preparative HPLCusing the standard acetonitrile/-water+0.1% TFA gradient and a VidacC-18 column. The pure sample had a M+1 ion at 704.3 in the mass spectrumand was homogenous by HPLC with a retention time of 24.12 minutes.

Example 144-(Piperidin-4-yl)carbonyl-1-{[2-(2-benzofuranoyl)amino]-4-[1-aminocarbonyl-2-(2-naphthyl)ethylamino]carbonylaminophenyl}homopiperazine

This compound was prepared by the method of Example 13 using2-benzofuran-carboxylic acid in the final coupling step to give thetitle compound with M+1 ion at 702.1 and retention time of 25.5 minutes.

Example 154-(Piperidin-4-yl)carbonyl-1-{[2-(2-benzofuranoyl)amino]-4-[1-aminocarbonyl-2-cyclohexylethylamino]carbonylaminophenyl}homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-cyclohexylalanine in the initial coupling step and2-benzofurancarboxylic acid for the final acylation step to give thetitle compound with M+1 ion at 658.3 and retention time of 25.64minutes.

Example 164-(Piperidin-4-yl)carbonyl-1-[2-[(2-benzofuranoyl)amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylaminophenyl]-homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-trans-4-aminomethylcyclohexanecarboxylic acid in the initialcoupling step and 2-benzofurancarboxylic acid for the final acylation togive the title compound with M+1 ion at 643.4 and retention time of21.84 minutes.

Example 17 4-(Methylaminomethyl)carbonyl-1-[2-[1(2-benzofuranoyl)amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylaminophenyl]homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-trans-4-aminomethylcyclohexanecarboxylic acid in the initialcoupling step and Boc-sarcosine for capping of the homopiperazine and2-benzofurancarboxylic acid for the final acylation to give the titlecompound with M+1 ion at 603.3 and retention time of 21.35 minutes.

Example 184-(Pyrrolidin-2-yl)carbonyl-1-[2-[(2-benzofuranoyl)amino]-4-[[(4-aminocarbonyl)cyclohexylmethylamino]carbonylamino]phenyl]homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-trans-4-aminomethylcyclohexanecarboxylic acid in the initialcoupling step, Boc-proline for capping of the homopiperazine and2-benzofurancarboxylic acid for the final acylation to give the titlecompound with M+1 ion at 629.3 and retention time of 22.12 minutes.

Example 194-(Piperidin-1-yl)-1-[2-[(2-benzofuranoyl)amino]-4-[[(4-amino-carbonyl)cyclohexylmethylamino]carbonylaminophenyl]-piperidine

This compound was prepared by the method of Example 13 usingFmoc-trans-4-aminomethylcyclohexanecarboxylic acid in the initialcoupling step, 4-(1-piperidyl)piperidine to displace the fluorine and2-benzofurancarboxylic acid for the final acylation to give the titlecompound with M+1 ion at 600.3 and retention time of 22.5 minutes.

Example 204-(Piperidin-4-yl)carbonyl-1-[[2-(4-phenylbutanoyl)amino]-4-[1-aminocarbonyl-2-cyclohexylethylamino]carbonylaminophenyl]homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-L-cyclohexylalanine in the initial coupling step andBoc-isonipecotic acid for capping of the homopiperazine to give thetitle compound with M+1 ion at 659.4 and retention time of 23.97minutes.

Example 21 [4-(Piperidin-4-yl)carbonyl-1-F[2-(4-phenylbutanoyl)amino]-4-[(1-aminocarbonyl-2-(naphth-2-yl))ethylamino]-carbonylaminophenyl]homopiperazine

This compound was prepared by the method of Example 13 usingFmoc-homophenylalanine in the initial coupling step to give the titlecompound with M+1 ion at 668.4 and retention time of 22.88 minutes.

Examples 22-25 describe the synthesis of sulfonamide compounds inaccordance with the compounds of the present invention.

Example 22N-(1-Aminocarbonyl-2-methylpropyl)-2-[(4-phenylmethyl)piperidin-1-yl]-5-[(2-pyrrolidinocarbonyl)amino]phenylsulfonamide

Following generally the procedures described above, commercialPolystyrene-RAM resin (0.74 mmol/g) (Rapp Polymere, Tubingen, Germany,0.25 g) was slurried in dichloromethane, washed with DMF and treated for30 minutes with a mixture of piperidine and DMF (1:1 v/v). The resin waswashed with DMF (5×), DCM (5×) and DMF (3×) and then coupled with 0.5mmol of Fmoc-(L)-valine, 1-hydroxybenzotriazole anddiisopropylcarbodiumide in 3 ml DMF over night. The resin was washedwith DMF (5×) and treated with piperidine/DMF again for 30 minutes.After washing with DMF (5×) and DCM (10×), the coupling with 0.5 mmol of2-fluoro-5-nitrophenylsulfonyl chloride in 2 ml DCM and 1 mmol oflutidine was carried out over night. The resin was washed with DCM (5×)and DMF (5×) and treated with 3 ml of a 0.5 molar solution of4-benzyl-piperidine in DMF for 24 hours at room temperature. Afterwashing with DMF (10×), the nitro group was reduced by shaking the resinwith 4 ml of a 0.5 molar solution of tin chloride in DMF/acetic acid 1:1for 72 hours. The resin was washed with DMF (5×), MeOH (5×), DCM (5×),DMF containing 5% of diisopropylethylamine (1×) and DMF (3×). The finalcoupling with 1 mmol of 2-pyrolidinecarboxylic acid,1-hydroxy-7-azabenzotriazole and diisopropylcarbodiimide in 3 ml DMF wasperformed over night. Following extensive washing of the resin with DMF,methanol and DCM and subsequent drying, it was cleaved with 3 ml of 95%trifluoroacetic acid. The TFA solution was evaporated and the residuewas combined with the washings of the resin with methanol. Evaporationyielded the crude title compound which was purified by preparative HPLCusing the standard acetonitrile/water+0.1% TFA gradient and a Vydac C-18column. The pure sample had a M+1 ion at 542.3 in the mass spectrum andwas homogenous by HPLC with a retention time of 26.7 minutes.

Example 23N-(1-Aminocarbonyl-2-methylpropyl)-2-[(4-phenylmethyl)piperidin-1-yl]-5-(4-piperdinocarbonyl)amino]phenylsulfonamide

This compound was prepared by the method of Example 22 using4-piperidinecarboxylic acid in the final coupling step to give the titlecompound with a M+1 ion at 556.3 in the mass spectrum and a HPLCretention time of 26.2 minutes.

Example 241-[2-[N-(2-Aminocarbonyl-3-methylbutyl)sulfonamido]-5-[2-cinnamoylamino]]phenyl-4-cyclohexylpiperazine

This compound was prepared by the method of Example 22 using1-cyclohexylpiperazine for displacement of the fluorine and cinnamicacid for the final acylation step to give the title compound with a M+1ion at 568.3 in the mass spectrum and a HPLC retention time of 24.63minutes.

Example 25N-[[(4-Aminocarbonyl)cyclohexylmethyl]amino]-[2-[(4-phenylmethyl)piperidin-1-yl]-5-[(2-pyrrolidinocarbonyl)-amino]phenyl]sulfonamide

This compound was prepared by the method of Example 22 using Fmocprotected trans-4-aminomethylcyclohexanecarboxylic acid for the firstcoupling reaction to give the title compound with a M+1 ion at 582.3 inthe mass spectrum and a HPLC retention time of 25.31 minutes.

Protein Tyrosine Kinase Activity

The compounds 1-25 above were assayed for activity with respect to theSrc protein tyrosine kinase by the fluorometric method described inMeasurement of the Protein Tyrosine Kinase Activity of c-Src UsingTime-Resolved Fluorometry of Europium Chelates, Braunwalder, A. F. etal., Analytical Biochemistry 238, 159-164 (1996), the disclosure ofwhich is incorporated herein by reference, using the materials andprocedures further specified below.

General Assay Method for Src-Kinase:

Materials:

-   Costar 384 Clear, non-treated, high-binding plates-   Sigma poly(Glu, Tyr) 4:1, ave. MW 35,000-   Src Kinase (p60^(C-Src))-   Sigma ATP (1.5 mM Stock Soln. in H₂O)-   MES Buffer: 30 mM MES (pH 6.8)-   10 mM MgCl₂-   MBI Buffer: (MES+0.4 mg/ml BSA+0.003% IGEPAL)-   Wallac Eu-labelled anti-phosphotyrosine Antibody (CR04-100)-   Coating Solution:-   22.5 μmnM Na₂CO₃ (pH 9.6)-   27.5 mM NaHCO₃-   0.9% NaCl-   Antibody Dilution Buffer: (MES+3% BSA)-   DELFIA® Wash Solution (TTBS):-   0.5 M NaC₁₋₂₀-   20 mM Tris (pH7.4)-   0.15% Tween 20-   DELFIA Enhancement Solution    Method:

The plates were coated with 0.1 mg/ml poly(Glu, Tyr) in CoatingSolution, 35 μl/well. It was let stand overnight at room temp. Theplates were then washed 3 times with MES (100 μl/wash).

Kinase Reaction Conditions:

Procedure (Listed in Order of Addition):

10 μl 200 μM A TP

80 nl 5 mM test compound in DMSO

10 μl 1:400 Src dilution in MBI

Final Reaction Conditions:

1:8000 Src Kinase

20 μM library compound (0.4% DMSO)

100 μM ATP

20 μL assay volume

15 min. at room temp.

The reaction was stopped by aspiration, and then washed 3 times with MES(100 μl/wash). 20 μl 0.4 ng/μl of antibody in Antibody Dilution Buffer(final=8 ng Ab/well) was added and then incubated for 30 mln. at RT. Theantibody solution was removed by aspiration and then washed 3 times with1× DELFIA Wash Solution. 20 μl DELFIA enhancement solution was added andthe plates were read on a Wallac Victor plate reader in time-resolvedfluorescence mode using 340 nm excitation and 615 nm emissionwavelengths.

The Src Kinase inhibitory activity of the compounds, given as IC50s(μM), are listed in Table 2.

TABLE 2 Compound No. Activity (μMol, Delfia assay. IC50) Example 1 22Example 2 23 Example 3 45 Example 4 18 Example 5 12.5 Example 6 14Example 7 13 Example 8 8.5 Example 9 17 Example 10 15.5 Example 11 11Example 12 18.5 Example 13 13 Example 14 2.75 Example 15 6.5 Example 1636 Example 17 37 Example 18 19 Example 19 22 Example 20 28 Example 21 22Example 22 14 Example 23 12 Example 24 42 Example 25 27.5

From these test results and the knowledge about the compounds describedin the references in the section “Background of the Invention”, it wouldbe apparent to the skilled artisan that the compounds of the inventionhave utility in treating conditions where selective inhibitory activityof an Src kinase is desirable. While the invention has been described indetail, modifications to illustrated embodiments within the spirit andscope of the present invention, set forth in the appended claims, willbe readily apparent to those of skill in the art.

1. A compound, including pharmaceutically acceptable salts or solvatesof said compound, said compound having the general structure shown inFormula II:

wherein R₁ is selected from the group consisting of H, straight chainC₁-C₆ alkyl; branched C₁-C₆ alkyl; —(CH₂)_(p)—Ar₁; and —(CH₂)_(p)—R₄,wherein p is 1 or 2; Ar₁ is phenyl or naphthyl optionally substitutedwith a straight chain or branched C¹-C⁶ alkyl group; and R₄ is C₅-C₇cycloalkyl; R₂ is selected from the group consisting of:

R₅ is selected from the group consisting of H, straight chain C₁-C₆alkyl; branched C₁-C₆ alkyl; R₃ is —(CH₂)_(q)—Ar₂ or —(CH═CH)-Phenyl,wherein q is an integer from 0 to 4; and Ar₂ is selected from the groupconsisting of:

X is:

Y is selected from the group consisting of:

with the proviso that when Y is any of the moieties:

then R₁ is H.
 2. The compound of claim 1, wherein R₁ is H.
 3. Thecompound of claim 1, wherein R₁ is a straight chain C₁-C₆ alkyl or abranched C₁-C₆ alkyl.
 4. The compound of claim 1, wherein R₁ is—(CH₂)_(p)—Ar₁, where p and Ar₁ are as defined in claim
 1. 5. Thecompound of claim 1, wherein R₁ is —(CH₂)_(p)—R₄, where p and R₄ are asdefined in claim
 1. 6. The compound of claim 1, wherein R₂ is:


7. The compound of claim 1, wherein R₁ is:


8. The compound of claim 1, wherein R₂ is:


9. The compound of claim 1, wherein R₃ is —(CH₂)_(q)—Ar₂, where q andAr₂ are as defined in claim
 1. 10. The compound of claim 1, wherein R₃is —(CH═CH)-Phenyl.
 11. The compound of claim 1, wherein Y is:


12. The compound of claim 1, wherein Y is:


13. The compound of claim 1, wherein Y is:


14. The compound of claim 1, wherein R₁ is isopropyl or isobutyl.
 15. Apharmaceutical composition comprising as an active ingredient a compoundof claim 1.